LOCUS       sequence-199053-        5604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  medium copy plasmid (pBR322 backbone) expressing the unstable GFP 
            variant GFP-OVA under control of the nitric oxide-inducible promoter
            hmpAp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5604)
  AUTHORS   Burton NA, Schurmann N, Casse O, Steeb AK, Claudi B, Zankl J, 
            Schmidt A, Bumann D
  TITLE     Disparate impact of oxidative host defenses determines the fate of 
            Salmonella during systemic infection in mice.
  JOURNAL   Cell Host Microbe. 2014 Jan 15;15(1):72-83. doi: 
            10.1016/j.chom.2013.12.006.
  PUBMED    24439899
REFERENCE   2  (bases 1 to 5604)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chom.2013.12"; journalName: "Cell Host Microbe"; date: 
            "2014-01-15- 15"; volume: "15"; issue: "1"; pages: "72-83"
COMMENT     Sequence Label: pNB22
FEATURES             Location/Qualifiers
     source          1..5604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..89
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     323..340
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    355..495
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(421..440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     581..603
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(597..788)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2246..2265
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             2830..3543
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2830..3543
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2860..2888)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3483..3503
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3668..3685)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3838..3924
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4016..4043
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4062..4153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4154..4222,4223..5014)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4372..4391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5185..5604,1..169)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
       61 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      121 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      181 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      241 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      301 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      361 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
      421 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
      481 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      541 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
      601 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
      661 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
      721 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
      781 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
      841 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
      901 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
      961 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     1021 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     1081 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     1141 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     1201 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     1261 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     1321 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     1381 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     1441 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     1501 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     1561 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     1621 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     1681 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     1741 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     1801 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     1861 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     1921 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     1981 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     2041 gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     2101 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     2161 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     2221 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     2281 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gagatcccct gacgtacttt
     2341 ttcctgctcc atagcctgcc acaattcggc atcataatcg gcaatgttca tttcacgctt
     2401 taacatccgc atctcctgac tcagctaaca ataaaatttg tgcctgaaaa ggcagtccgt
     2461 gggacttcgg ccaacagtat aaccgaatca ttcttcgata acaggtcttg acaaaggttt
     2521 ttacgcaaac gattacctat gcgtcagata agggtttcct gaacgagagt ctgacgaatt
     2581 tcaacggatt tcttttcagc tttgtgatgc agatttttca cgttgttacc tccataacgt
     2641 aaagcagaga agatccattt acaacgcaag ggtattttta taagatgcat ttgatataca
     2701 tcattagatt ttcacataaa ggaagcacgt atgcttgacg cacaaaccat cgctacagta
     2761 aaggccacca ttcccctgct ggttgaaaca ggaccgaaac tgatctagat ttaagaagga
     2821 gatatacata tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     2881 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     2941 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     3001 ccaacacttg tcactacttt cgcgtatggt cttcaatgct ttgcgagata cccagatcat
     3061 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
     3121 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     3181 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     3241 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     3301 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     3361 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     3421 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     3481 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     3541 aaagaatctc tgaaaatctc tcaggctgtt cacgctgctc acgctgaaat caacgaagct
     3601 ggtcgtgaag tagtaggtta actgcagcca agcttggctg ttttggcgga tgagagaaga
     3661 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     3721 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     3781 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     3841 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3901 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     3961 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     4021 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     4081 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4141 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4201 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4261 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4321 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4381 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     4441 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4501 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4561 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4621 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4681 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4741 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4801 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4861 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4921 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4981 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5041 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5101 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5161 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5221 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5281 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5341 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5401 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5461 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5521 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5581 agaaagcgcc acgcttcccg aagg
//
